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The Phylogenetic Relationship Revealed Three New Wood-Inhabiting Fungal Species from Genus Trechispora

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The Phylogenetic Relationship Revealed Three New Wood-Inhabiting Fungal Species from Genus Trechispora fmicb-12-650195 March 11, 2021 Time: 17:8 # 1 ORIGINAL RESEARCH published: 17 March 2021 doi: 10.3389/fmicb.2021.650195 The Phylogenetic Relationship Revealed Three New Wood-Inhabiting Fungal Species From Genus Trechispora Wei Zhao1,2 and Chang-Lin Zhao1,2* 1 Key Laboratory for Forest Resources Conservation and Utilization in the Southwest Mountains of China, Ministry of Education, Southwest Forestry University, Kunming, China, 2 College of Biodiversity Conservation, Southwest Forestry University, Kunming, China Wood-inhabiting fungi play a significant role in wood degradation and the cycle of matter in the ecological system. In the present study, three new wood-inhabiting fungal species, Trechispora bambusicola, Trechispora fimbriata, and Trechispora fissurata spp. nov., are Edited by: Dhanushka Nadeeshan nested in Trechispora, which are proposed based on a combination of morphological Wanasinghe, features and molecular evidence. Sequences of internal transcribed spacer (ITS) Key Laboratory for Plant Diversity and Biogeography of East Asia, and large subunit (nLSU) regions of the studied samples were generated, and the Kunming Institute of Botany, Chinese phylogenetic analyses were performed with maximum likelihood, maximum parsimony, Academy of Sciences, China and Bayesian inference methods. The phylogenetic analyses inferred from ITS showed Reviewed by: that T. bambusicola was sister to Trechispora stevensonii, T. fimbriata grouped Baokai Cui, Beijing Forestry University, China with Trechispora nivea, and T. fissurata grouped with Trechispora echinospora. The Yu-Cheng Dai, phylogenetic tree based on ITS C nLSU sequences demonstrated that T. bambusicola Beijing Forestry University, China formed a single lineage and then grouped with Trechispora rigida and T. stevensonii. *Correspondence: Chang-Lin Zhao T. fimbriata was sister to T. nivea. T. fissurata grouped with Trechispora thelephora. [email protected] Keywords: Hydnodontaceae, phylogeny, taxonomy, wood-inhabiting fungi, Yunnan Province Specialty section: This article was submitted to Evolutionary and Genomic INTRODUCTION Microbiology, a section of the journal Trechispora P. Karst. (Hydnodontaceae, Trechisporales) was typified with Trechispora onusta Frontiers in Microbiology P. Karst. (Karsten, 1890). It is characterized by the resupinate to effused basidiomata with Received: 08 January 2021 smooth to hydnoid to poroid hymenophore, a monomitic or dimitic hyphal structure Accepted: 01 February 2021 with clamped generative hyphae having typical ampullaceous septa, and short cylindric Published: 17 March 2021 Citation: Zhao W and Zhao C-L (2021) The Abbreviations: ITS, internal transcribed spacer; nLSU, large subunit; SWFC, herbarium of Southwest Forestry University, Kunming, China; KOH, 5% potassium hydroxide; CB, Cotton Blue; CB−, acyanophilous; IKI, Melzer’s reagent; IKI−, both Phylogenetic Relationship Revealed inamyloid and indextrinoid; L, mean spore length (arithmetic average for all spores); W, mean spore width (arithmetic Three New Wood-Inhabiting Fungal average for all spores); Q, variation in the L/W ratios between The studied specimens, n (a/b), number of spores (a) measured Species From Genus Trechispora. from given number (b) of specimens, spore measurements do not include ornamentation; CTAB, cetyltrimethylammonium Front. Microbiol. 12:650195. bromide; DNA, deoxyribonucleic acid; PCR, polymerase chain reaction; MP, maximum parsimony; ML, maximum doi: 10.3389/fmicb.2021.650195 likelihood; BI, Bayesian inference; TBR, tree-bisection reconnection. Frontiers in Microbiology| www.frontiersin.org 1 March 2021| Volume 12| Article 650195 fmicb-12-650195 March 11, 2021 Time: 17:8 # 2 Zhao and Zhao Three Potential New Ectomycorrhizal Species basidia and smooth to verrucose or aculeate basidiospores based on field notes. Color terms follow Petersen(1996). (Karsten, 1890; Bernicchia and Gorjón, 2010). About 49 species Micromorphological data were obtained from the dried are currently known in Trechispora worldwide (Liberta, 1966, specimens and observed under a light microscope following 1973; Larsson, 1994, 1995, 1996; Ryvarden, 2002; Trichiès and Dai(2012). The following abbreviations were used: KOH = 5% Schultheis, 2002; Miettinen and Larsson, 2006; Ordynets et al., potassium hydroxide, CB = Cotton Blue, CB− = acyanophilous, 2015; Xu et al., 2019) and Index Fungorum1 and MycoBank2. IKI = Melzer’s reagent, IKI− = both inamyloid and indextrinoid, Larsson(2007) addressed the classification of corticioid fungi L = mean spore length (arithmetic average for all spores), and revealed that Trechispora farinacea (Pers.) Liberta grouped W = mean spore width (arithmetic average for all spores), with Trechispora hymenocystis (Berk. and Broome) K.H. Larss., Q = variation in the L/W ratios between the studied specimens, n in which both species nested within the family Hydnodontaceae (a/b) = number of spores (a) measured from given number (b) of Jülich. Based on the large subunit nuclear ribosomal RNA gene specimens, spore measurements do not include ornamentation. (nLSU) datasets, Albee-Scott and Kropp(2010) supported to transfer Hydnodon thelephorus (Lév.) Banker to Trechispora as Molecular Phylogeny Trechispora thelephora (Lév.) Ryvarden. The order Trechisporales Cetyltrimethylammonium bromide (CTAB) rapid plant genome was studied employing the internal transcribed spacer (ITS) extraction kit-DN14 (Aidlab Biotechnologies Co., Ltd., Beijing, and nLSU regions, in which it suggested that Porpomyces China) was used to obtain genomic deoxyribonucleic acid Jülich, Sistotremastrum J. Erikss., Subulicystidium Parmasto, (DNA) from dried specimens, according to the manufacturer’s and Trechispora belonged to a highly supported clade and instructions following Zhao and Wu(2017). ITS region was Trechispora belongs to Hydnodontaceae and was closely related amplified with primer pair ITS5 and ITS4 (White et al., 1990). to Brevicellicium K.H. Larss. and Hjortstam (Telleria et al., Nuclear LSU region was amplified with primer pair LR0R and 2013). A phylogenetic study of Trechispora was addressed and LR73. The polymerase chain reaction (PCR) procedures for ITS demonstrated that Trechispora cyatheae Ordynets, Langer and and nLSU following Zhao and Wu(2017). The PCR products K.H. Larss. and Trechispora echinocristallina Ordynets, Langer were purified and directly sequenced at Kunming Tsingke and K.H. Larss. clustered into Trechispora as new members, Biological Technology Limited Company, Kunming, Yunnan inferred from the combined data of the ITS and LSU datasets Province, China. All newly generated sequences were deposited (Ordynets et al., 2015). The phylogeny of Trechisporales was at GenBank (Table 1). inferred from a combined dataset of ITS-nLSU sequences and Sequencher 4.6 (GeneCodes, Ann Arbor, United States) was showed that Porpomyces, Scytinopogon Singer, and Trechispora used to edit the DNA sequence. Sequences were aligned in grouped together and nested within family Hydnodontaceae (Liu MAFFT 74 using the “G-INS-I” strategy and manually adjusted et al., 2019). Phylogram generated from analysis of ITS sequence in BioEdit (Hall, 1999). The sequence alignment was deposited dataset of Trechispora showed that Trechispora echinospora in TreeBase (submission ID 25879). Sequences of Fibrodontia Telleria was sister to the clade formed by Trechispora araneosa alba Yurchenko and Sheng H. Wu and Fibrodontia gossypina (Hohn. and Litsch.) K.H. Larss., T. farinacea, T. hymenocystis, Parmasto retrieved from GenBank were used as an outgroup in and Trechispora mollusca (Pers.) Liberta with a low support the ITS C nLSU analyses by following Ordynets et al.(2015). (Phookamsak et al., 2019). The ITS C nLSU dataset comprised Maximum parsimony (MP) analyses were applied to the 22 species and revealed that Trechispora yunnanensis C.L. Zhao ITS C nLSU dataset sequences. Approaches to phylogenetic formed a monophyletic lineage within Trechispora and was analysis followed Zhao and Wu(2017), and the tree construction closely related to Trechispora byssinella (Bourdot) Liberta and procedure was performed in PAUP∗ version 4.0b10 (Swofford, Trechispora laevis K.H. Larss. (Xu et al., 2019). 2002). All characters were equally weighted and gaps were During the studies on wood-inhabiting fungi in southern treated as missing data. Trees were inferred using the heuristic China, three species of Trechispora could not be assigned search option with tree-bisection reconnection (TBR) branch to any described species. Obtaining sequences from the new swapping and 1000 random sequence additions. Max-trees were taxa, the authors examine taxonomy and phylogeny of three set to 5000, branches of zero length were collapsed, and all new species within the genus Trechispora, based on the ITS parsimonious trees were saved. Clade robustness was assessed and nLSU sequences. using a bootstrap (BT) analysis with 1000 replicates (Felsenstein, 1985). Descriptive tree statistics tree length (TL), consistency index (CI), retention index (RI), rescaled consistency index (RC), MATERIALS AND METHODS and homoplasy index (HI) were calculated for each Maximum Parsimonious Tree generated. Datamatrix was also analyzed Morphology using maximum likelihood (ML) approach with RAxML-HPC2 The studied specimens are deposited at the herbarium of through the Cipres Science Gateway with GTR C I C G molecular Southwest Forestry University (SWFC), Kunming, Yunnan evolution model5 (Miller et al., 2009). Branch support (BS) for Province, China. Macromorphological descriptions
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